Abstract
Enterovirus 71 (EV71) is the main causative agent of severe hand-foot-mouth disease. EV71 affects countries mainly in the Asia-Pacific region, which makes it unattractive for pharmaceutical companies to develop drugs or vaccine to combat EV71 infection. However, development of these drugs and vaccines is vital to protect younger generations. This study aims to develop a specific monoclonal antibody (mAb) to EV71 using a plant platform, which is a cost-effective and scalable production technology. A previous report showed that D5, a murine anti-EV71 mAb, binds to VP1 protein of EV71, potently neutralizes EV71 in vitro, and effectively protects mice against EV71 infection. Herein, plant-produced chimeric D5 (cD5) mAb, variable regions of murine D5 antibody linked with constant regions of human IgG1, was transiently expressed in Nicotiana benthamiana using geminiviral vectors. The antibody was expressed at high levels within six days of infiltration. Plant-produced cD5 retained its in vitro high-affinity binding and neutralizing activity against EV71. Furthermore, a single dose (10 µg/g body weight) of plant-produced cD5 mAb offered 100% protection against infection in mice after a lethal EV71 challenge. Therefore, our results showed that plant-produced anti-EV71 mAb is an effective, safe, and affordable therapeutic option against EV71 infection.
Highlights
Hand-foot-mouth disease (HFMD) mainly affects children under five-years-old
To construct chimeric monoclonal antibody (mAb), variable regions of murine D5 antibody were linked with constant regions of human IgG1 heavy chain (HC) and kappa light chain (LC) and separately cloned into a geminiviral vector, resulting in pBYD5-HC and pBYD5-LC (Figure 1)
The leaves infiltrated with pBYD5-HC and pBYD5-LC showed necrosis on the sixth day after infiltration (Figure 2A)
Summary
Hand-foot-mouth disease (HFMD) mainly affects children under five-years-old. HFMD is of considerable public health concern in the Asia-Pacific region because of the economic burden in the affected countries. In addition to hospital expenditure, loss of time from work is an additional cost, because families have to care for the sick children away from school [1]. Considering the economic burden of HFMD, the development of HFMD vaccine and a therapeutic drug is required urgently. HFMD is caused by several viruses such as enterovirus 71 (EV71) and coxsackieviruses [2]. EV71 causes severe central nervous system or cardiovascular complications, leading to the death of infected patients [3,4]. Various types of EV71 vaccines have been developed previously, such as inactivated whole-virus vaccines [5,6,7,8,9], live attenuated vaccines [10,11], recombinant vaccines [12,13,14,15], and peptide
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